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@PHDTHESIS{Ning:17113,
author = {Ning, Jing},
title = {{A}pplication of functional gene arrays for monitoring
influences of plant/seasons on bacterial functions and
community structures in constructed wetland ({B}itterfeld,
{G}ermany)},
volume = {114},
issn = {1866-1793},
school = {RWTH Aachen},
type = {Dr. (Univ.)},
address = {Jülich},
publisher = {Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag},
reportid = {PreJuSER-17113},
isbn = {978-3-89336-724},
series = {Schriften des Forschungszentrums Jülich : Energie $\&$
Umwelt / Energy $\&$ Environment},
pages = {XIV, 157 S.},
year = {2011},
note = {Record converted from VDB: 12.11.2012; RWTH Aachen, Diss.,
2011},
abstract = {With the increasing application of constructed wetlands
(CWs) around the world to improve water quality, it becomes
more and more important to gain details about microbial
ecology in this “black box”. In order to better
understand general relationships among bacterial community
structures, functions and environmental influencing factors
in CWs, soil samples taken from the planted and the
unplanted CWs in Bitterfeld (Germany), as well as those
taken in warm- and cold-seasons, were compared in this study
by using functional gene arrays (the GeoChip). In addition,
other assessment methods, such as chemical analyses,
fluorescent microscopic enumeration and PCRbased DGGE
($\underline{D}$enaturing $\underline{G}$radient
$\underline{G}$el $\underline{E}$lectrophoresis) were also
applied to complete the conclusions obtained by using the
GeoChip. In general, this study consisted of two parts. The
first part focused on evaluations of suitable DNA
preparation procedures for functional gene arrays. The whole
genome amplification was taken out of the primary plan due
to biases in proportions of bacterial populations in DGGE
band patterns, which were observed not only among the
replications of the same DNA template but also between the
products with and without this treatment. While evaluating
procedure of DNA purification, the gel filtration column
failed to purify DNA extracts of our samples, probably due
to high amount of humic substances in the extracts. Among
the three DNA extraction kits compared in this study (e.g.
Bio101 FastDNA Spin Kit for Soil, UltraClean Soil DNA
Isolation Kit and PowerSoil DNA Isolation Kit), extracts of
the Bio101 were proved to be more suitable for the DGGE
analysis with respect to their purity, yield and
representativeness, while those of the PowerSoil showed
better results in the microarray analysis. In addition,
basing on the DNA extracts revealing different quantity and
purity indices (A260/280, A260/230 and A320), influences of
these indices on subsequent assessment methods of microbial
communities (e.g. PCR-based DGGE and functional gene arrays)
were also investigated. It was concluded that the success in
PCR performance and consequently also in the DGGE analysis
is more affected by the A260/280 and A320 values than by the
ratio A260/230, and conditionally by the DNA yield.
Moreover, the DGGE band pattern could also be affected by
the preferential extraction due to chemical agents applied
in the extraction. However, unlike the DGGE analysis, DNA
microarrays were more affected by the A260/230 and A320
values. Thus, the primary goal of DNA extraction to obtain
DNA extracts with the highest purity is not accurate enough
any more in the environmental studies applying various
analyzing methods. The performance of DNA extraction should
vary with the subsequent [...]},
cin = {IBG-3},
cid = {I:(DE-Juel1)IBG-3-20101118},
pnm = {Terrestrische Umwelt},
pid = {G:(DE-Juel1)FUEK407},
typ = {PUB:(DE-HGF)11 / PUB:(DE-HGF)3},
url = {https://juser.fz-juelich.de/record/17113},
}